Biofunctionalization of Poly(acrylamide) Gels.

Engineering novel biomaterials that mimic closer in vivo scenarios requires the simple and quantitative incorporation of multiple instructive signals to gain a higher level of control and complexity at the cell-matrix interface. Poly(acrylamide) (PAAm) gels are very popular among biology labs as 2D model substrates with defined biochemical and mechanical properties. These gels are cost effective, easy to prepare, reproducible, and available in a wide range of stiffness. However, their functionalization with bioactive ligands (cell adhesive proteins or peptides, growth factors, etc.) in a controlled and functional fashion is not trivial; therefore reproducible and trustable protocols are needed. Amine or thiol groups are ubiquitous in natural or synthetic peptides, proteins, and dyes, and hence routinely used as handles for their immobilization on biomaterials.We describe here the preparation of mechanically defined (0.5-100 kPa, range that approximates the stiffness of most tissues in nature), thin PAAm-based hydrogels supported on a glass substrate and covalently functionalized with amine- or thiol-containing bioligands via simple, robust, and effective protocols.